Project Summary Synthetic, or designer, drugs represent a serious public health risk. Compared to the traditional drugs for which they are sold as substitutes, synthetic drugs are often more potent and can result in life-threatening medical complications. One of the difficulties in preventing complications from synthetic drugs is that they go undetected by routine drug screens used in hospitals. Clinicians are therefore unaware of what the patient took. Moreover, determining when new synthetic drugs hit the street and whether there are complications related to their abuse is delayed. While samples can be sent to reference laboratories for testing, the results often do not come back for days or weeks making the tests of little help for the involved clinicians and are therefore rarely sent. Mass spectrometry is an attractive option for performing synthetic drug screens because it is sensitive, selective, and is a platform technology that can adapted relatively quickly as new agents emerge. The use of mass spectrometry for this purpose at or near the point of care is limited, however, due to the complexity of the analyses. The focus of this proposal is to simplify the entire analytical workflow required to perform mass spectrometric assays in order to make the use of MS assays more feasible for rapid screening of synthetic drugs. The core technology in the proposal is paper spray mass spectrometry, a direct analysis method in which biofluids samples such as blood can be analyzed directly without sample preparation. All of the components needed to perform the extraction and ionization of analytes from complex matrices can be contained within an inexpensive disposable cartridge. No sample preparation is performed, no solvent waste is generated, and no carry-over or clogging occur. These advantages make paper spray MS an attractive option for synthetic drug screening. The sensitivity of paper spray MS needs to be improved by about an order of magnitude to make it adequate for screening synthetic cannabinoids and fentanyl derivatives, however. In this proposal, we aim to develop and test a disposable paper spray cartridge that can dramatically improve the sensitivity of paper spray MS without increasing the complexity of the analysis. The general approach is to perform analyte preconcentration and separation from the sample matrix automatically on an inexpensive disposable cartridge, one requiring no special device or human action. We will also begin testing these methods on blood samples of patients presenting to the Emergency Department who are suspected to have used synthetic drug.